KR20150028308A - Peptide nanoparticles and uses therefor - Google Patents

Abstract

The present invention provides nanoparticle compositions comprising one or more peptides. The present invention transdermally delivers such peptides without peptide modification for use in physical or chemical rubbing or rupturing of the skin.

Description

Peptide nanoparticles and uses thereof {PEPTIDE NANOPARTICLES AND USES THEREFOR}

Related application

This application is filed under 35 U.S.C. §119(e) U.S. Provisional patent application, U.S.S.N. 60/872,206, (filed on December 1, 2006) (hereinafter referred to as the '206 application') is claimed as priority, and the full text of the 206 application is attached as a reference.

Peptides have been found to provide cosmetically and therapeutically beneficial effects on the skin. In experimental models, short peptides (up to 30 amino acids in length) have been found to promote collagen growth on the extracellular matrix of the skin, and may improve the appearance of the skin and the treatment of damaged skin (Katayama, et ah, 1993, J. Biol. Chem., 268:9941; incorporated herein by reference).

Modified peptides have been shown to reduce wrinkle formation through the regulation of enzymes that affect muscle contraction of the muscles under the skin involved in wrinkle formation (Lupo, 2005, Dermatol. Surg., 31:832; incorporated herein by reference).

However, a major problem in obtaining the potential cosmetic and therapeutic effects of these peptides in humans is the transport of the peptides transdermally through the outer skin barrier (stratum corneum) to the site of biological action, such as the extracellular matrix or underlying muscles. (Robinson, et ah, 2005, International J. Cosmetic Science 27: 155; incorporated herein by reference). For transdermal delivery of peptides in humans, the peptides must be modified by adding chemical moieties including, but not limited to, acetyl and/or palmitoyl groups (Robinson, et al, supra). Such chemical modifications are expensive and time consuming, and have disadvantages that negatively affect the commercial production of products containing these peptides. Chemical modification of the peptide can also reduce the biological activity by diminishing the effect by reducing the ability to bind to cellular receptors at the biologically active site (eg spatial interference). Peptides that are less biologically effective will be less effective for cosmetic or therapeutic purposes.

Peptides that are less biologically effective would have to be administered in higher dosages to obtain their desired biological effect (if any), which would be a cost disadvantage for the commercial production of the product.

Summary of the invention

The present invention describes nanoparticles in which short, unmodified peptides (2 to 30 amino acids in length), which are biologically active ingredients, are bound to skin (epidermis and dermis), subcutaneous tissue (including adipose tissue), and lateral muscles.

The nanoparticles of the present invention can be provided to the skin of an individual. In some embodiments, the nanoparticles are transdermally transported of the peptide bound to the subject.

The nanoparticles of the present invention may be mixed with one or more excipients or provided to the skin as simple suspensions or dispersions, and emollients, nutritional lotions, cleansing lotions, cleansing creams, skin milk, emollient lotions, massage creams, emollient creams. , Makeup base, lipstick, face pack or face gel, cleaner composition (e.g., shampoo, conditioner, body cleanser, hair tonic, and soap), skin composition (e.g. lotion, ointment, gel, cream, patch, Spray), etc.

Accordingly, the present invention provides systems and compositions for transdermal transport of unmodified peptides. Among the many advantages of the present invention is that the peptide can be delivered without injection, and additionally can be delivered without mechanical or chemical friction to the skin or deformation of the skin. Additional advantages include the ability to use unmodified peptides, simplifying and reducing the cost of production of the cosmetic and/or pharmaceutical preparations of the present invention, preserving the biological activity of the peptides.

Justice

Abrasion: As used herein, friction refers to anything that alters, ruptures, removes, or destroys the upper layer of skin. In some embodiments, friction refers to a mechanical means of altering, rupturing, removing or destroying the upper layer of skin. In some embodiments, friction refers to a chemical means of altering, rupturing, removing or destroying the upper layer of skin. Exfoliating agents, fine particles (eg magnesium or aluminum particles), acids (eg alpha-hydroxy acids or beta-hydroxy acids), alcohols may cause friction, to name a few. In general, Donovan (e.g., U.S. Patent Publication 2004/009180; 2005/175636; PCT Publication WO 04/06954; all incorporated by reference), and Graham (e.g., U.S. Patent 6,939,852; U.S. Patent Publication 2006/ The penetration reinforcement described in 093624; this is attached as a reference) is considered to be the cause of friction. Of course, one of ordinary skill in the art will recognize that at a certain concentration, certain substances may cause friction, or when certain substances are associated with one or more other substances, they may cause friction, but under different conditions they do not cause friction. Therefore, whether a particular substance becomes a “friction substance” depends on the content. One of skill in the art would be able to immediately assess friction by observing redness or skin irritation, or by observing a histological examination of the skin showing deformation, rupture, removal or erosion of the stratum corneum.

Amino Acid: As used herein, “amino acid” refers to any compound and/or substance that is bound to a polypeptide chain in a broad sense. In some embodiments, the amino acid has the general structure H ₂ NC(H)(R)-COOH. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a synthetic amino acid; In some embodiments, the amino acid is a D-amino acid; In some embodiments, the amino acid is an L-amino acid.

Standard amino acids (“Standard amino acids”) refer to any of the 20 standard L-amino acids found in naturally occurring peptides. Nonstandard amino acids (“Nonstandard amino acids”) refer to any amino acid other than standard amino acids, whether obtained synthetically or prepared from natural sources. Amino acids containing carboxy and/or amino-terminal amino acids in the peptide are methylated, amidated, acetylated, and/or modified by substitution with any chemical group capable of changing the circulating half-life without adversely affecting the activity of the peptide. Can be. However, as mentioned herein, the present invention specifically relates to “unmodified peptides”, which refer to peptides that have not been chemically modified to effect or achieve transdermal transport. Amino acids can participate in disulfide bonds. “Amino acid” is used interchangeably with “amino acid residue” and refers to free amino acids and/or amino acid residues of peptides. Whether the term used herein refers to a free amino acid or an amino acid residue of a peptide can be confirmed by content.

Animal: As used herein, “animal” refers to any animal in the animal kingdom. In some embodiments, “animal” refers to a person at any stage of development. In some embodiments, “animal” refers to a mammal (eg, rodent, mouse, field mouse, rabbit, monkey, dog, cat, sheep, cow, primate, and/or pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, or worms. In some embodiments, the animal may be a transgenic animal, a genetically engineered animal, and/or a clone.

Approximately: As used herein, the terms “approximately” and “about” refer to values that are similar to the standard reference values as applied to one or more values of possession. In certain embodiments, “approximately” or “about” means 25% or 20% of the stated numerical value, unless otherwise stated (except for cases exceeding 100% of the expected value). , 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3 It refers to a range of values that fall within the range of %, 2%, 1%, or less.

Biologically active agent: As referred to herein, "biologically active substance" refers to any substance that is active in living systems and/or organisms.

For example, a substance having a biological effect on the organism is considered to be biologically active when administered to the organism. In certain embodiments, where the protein or polypeptide is biologically active, the portion of the protein or polypeptide that shares at least one biological activity of the protein or polypeptide is also generally referred to as a “biologically active” moiety.

Botulinum toxin: As used herein, botulinum toxin refers to any neurotoxin made by Clostrinium botulinum. Unless otherwise stated, the term includes fragments or portions (eg, light and/or heavy chains) of neurotoxins (eg, muscle relaxation activity) that retain appropriate activity. The botulinum toxin referred to herein includes botlinium toxin serotypes A, B, C, D, E, F, and G. As mentioned herein, botlinium toxins include botulinum toxin complexes (e.g., 300, 600, 900 kD complexes) as well as purified (e.g., isolated) botlinium toxins (e.g. , About 150kD).

"Purified botlinium toxin" is defined as a botulinum toxin isolated or substantially isolated from other proteins, including the botlinium toxin complex protein. The purified toxin has a purity of 95% or more, and preferably a purity of 99% or more. Those of skill in the art will appreciate that the present invention is not limited to any particular source of botulinum toxin. For example, the botulinum toxin that can be used in the present invention is isolated from Clostridium botulinum , chemically synthesized, or recombinant (e.g., an organism or host cell other than Clostridium botulinum). In) can be.

“Characteristic portion”: As mentioned herein, “characteristic portion” of a substance in a broad sense means that it shares at least one functional characteristic and/or some degree of sequence or structural identity of the relevant intrinsic substance. do. For example, a “characteristic portion” of a protein or polypeptide is one comprising a contiguous stretch amino acid or a collection of contiguous stretch amino acids that are metric of the protein or polypeptide. In some embodiments, such continuous stretches will contain at least 2, 5, 10, 15, 20 or more amino acids. In general, the characteristic portion shares at least one functional characteristic of the relevant native protein, in addition to the sequence identity specified above. In some embodiments, the characteristic moiety is biologically active.

Hydrophilic: As described herein, a “hydrophilic” material is a material that is soluble in polar solvents. In some embodiments, the hydrophilic material can temporarily bind with a polar solvent. In some embodiments, the hydrophilic material binds to the polar solvent through hydrogen bonding. In some embodiments, the polar solvent is water. In some embodiments, the hydrophilic material is ionic.

In some embodiments, the hydrophilic material is non-ionic. In some embodiments, the hydrophilic material can be dissolved directly in water, polar solvents, or hydrophilic solvents rather than oils. In some embodiments, the hydrophilic material may be less soluble in oils, non-polar solvents or hydrophobic solvents than water, polar solvents, or hydrophilic solvents.

In some embodiments, the material is hydrophilic relative to other materials because it is more soluble in water, polar solvents, or hydrophilic solvents than other materials. In some embodiments, the material is hydrophilic compared to other materials because it is less soluble in oils, non-polar solvents, or hydrophobic solvents than other materials.

Hydrophobic: As used herein, a “hydrophobic” material is a material that is soluble in a non-polar solvent. In some embodiments, the hydrophobic material is rejected from polar solvents. In some embodiments, the polar solvent is water. In some embodiments, the hydrophobic material is non-polar. In some embodiments, the hydrophobic material may be more soluble in oils, non-polar solvents or hydrophobic solvents than water, polar solvents, or hydrophilic solvents. In some embodiments, the hydrophobic material may be less soluble in water, polar solvents, or hydrophilic solvents than in oils, non-polar solvents, or hydrophobic solvents. In some embodiments, the material is hydrophobic relative to other materials because it is more soluble in oils, non-polar solvents, or hydrophobic solvents than other materials. In some embodiments, the material is hydrophobic to other materials because it is less soluble in water, polar solvents, or hydrophilic solvents than other materials.

In conjunction with: As used herein, “delivered in conjunction with” refers to the joint transport of two or more substances or drugs. In particular, according to the present invention, this term is used to refer to the transport of the nanoparticles and/or nanoparticle compositions of the present invention and biologically active substances.

The substance or drug is transported with the nanoparticles when combined with the nanoparticles and/or nanoparticle composition; The substance or drug is entrapped or completely enclosed by the nanoparticles; The substance or drug is embedded within a nanoparticle micellar membrane; The substance or drug is associated with the outer surface of the nanoparticle micelle membrane. The substance or drug carried with the nanoparticles and/or nanoparticle composition may or may not be covalently linked to the nanoparticle and/or nanoparticle composition. The substance or drug to be transported with the nanoparticles and/or nanoparticle composition of the present invention may or may not be attached to the nanoparticle and/or nanoparticle composition by adsorption force.

Isolated: As used herein, the term "isolated" is (1) separated from at least a portion of the components associated with it when it was first made (either in its natural state or in a laboratory setting), and /Or (2) refers to substances and/or entities produced, prepared and/or manufactured by human hands. Separated substances and/or entities are at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90 of the other components to which they are originally associated. Can be separated from %, or more. In some embodiments, the isolated substances and/or entities have 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% purity.

Microfluidized: As used herein, “microfluidized” means exposure to high shear forces. In some embodiments, exposure to such high shear forces is achieved by exposure to high pressure; In some embodiments, such high pressure ranges from about 15,000 to about 26,000 psi. In some embodiments, such exposure to high shear forces is achieved by cavitation. In some embodiments, such exposure to high shear forces is achieved by passing the sample through an instrument such as a Microfluidizer (Microfluidics Corporation/MFIC Corporation) or other similar device that can be used to make a homogeneous nanoparticle composition. In some embodiments of the present invention, the sample is microfluidized through exposure to high shear forces for less than about 10 minutes. In some embodiments, the time is less than about 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, or 1 minute. In some embodiments, the time is in the range of about 1-2 minutes. In some embodiments, the time is about 30 seconds. In some embodiments of the invention, the sample is “microfluidized” through a single exposure to high shear forces; This specific example is called “single pass” microfluidization.

Nanoparticle: As used herein, “nanoparticle” refers to any particle with a diameter of less than 1000 nm. In some embodiments, nanoparticles have a diameter of less than 300 nm, as defined by the National Science Foundation. In some embodiments, the nanoparticles are less than 100 nm in diameter as defined by the National Institutes of Health. In some embodiments, the nanoparticles are composed of trapping compartments separated from the bulk solution by micelle membranes as micelles. A “micellar membrane” is an amphiphilic entity that encloses a space or septum and is agglomerated to capture (eg, define a lumen).

Nanoparticle composition: As used herein, “nanoparticle composition” refers to any material comprising at least one nanoparticle. In some embodiments, the nanoparticle composition is a homogeneous collection of nanoparticles. In some embodiments, the nanoparticle composition is a dispersion or emulsion. In general, dispersions or emulsions are formed by combining at least two unmixed substances.

Hydrophilic emulsified ("oil-in-water") dispersions are those in which oily particles (or hydrophobic or non-polar) are dispersed in an aqueous dispersion medium. Lipophilic emulsification ("water-in-oil") is the dispersion of water-soluble (or hydrophilic or polar) particles in an oily dispersion medium. Those skilled in the art will understand that dispersions can be formed from any two immiscible media, but are not strictly limited to water-soluble and oily media combinations. The “dispersion medium” is widely applied as any dispersion medium, although it is commonly referred to in the “aqueous” and “oily” categories. In some embodiments, the nanoparticle composition is a nanoemulsion. In some embodiments, the nanoparticle composition is composed of micelles In some specific embodiments, the nanoparticle composition is PCT/US07/-------, filed on Nov. Entity nanoparticles are composed of amphoteric entity nanoparticles as described in'entity nanoparticles' (attached by reference). In some embodiments, the nanoparticle composition is stable In some embodiments, the nanoparticle composition includes nanoparticles and It contains one or more biologically active substances to be carried together.

Nutraceutical: As referred to herein, “nutraceutical” refers to any substance that is considered to provide a medical, health or biological benefit. In some embodiments, the nutraceutical will be able to prevent disease. In some embodiments, nutraceuticals provide basic nutritional value. In some embodiments, the nutraceutical is a food product or part of a food product. In some embodiments, the nutraceutical material may be isolated nutrients, dietary supplements, vitamins, minerals, herbs, supplements, therapeutic foods, genetically engineered foods and processed foods. Functional foods are also referred to as “phytochemical foods” or “functional foods”.

Premix: As used herein, “premix” refers to any combination of ingredients used to make a nanoparticle composition according to the present invention. For example, a premix is any collection of components that, when exposed to high shear forces, produce nanoparticles according to the invention. In some embodiments, the premix includes two or more immiscible solvents. In some embodiments, a premix includes components that self-assemble into nanoparticles. In some embodiments, the nanoparticle composition in the premix is as described in PCT/US07/-------,'Bilateral Entity Nanoparticles' filed on November 30, 2007 (Attached by reference ), including one or more amphoteric entity nanoparticles. In some embodiments, the premix includes one or more unmodified peptides; In some embodiments, the premix contains at least one biologically active substance. In some embodiments, the premix is perturbed, mixed or agitated; In some embodiments, the premix is perturbed, mixed, or agitated prior to receiving a high shear force. In some embodiments, the premix consists of at least one solubilizing component (eg, at least one component in solution); In some embodiments, the premix is subjected to a high shear force after solubilization.

Pure: As used, substances and/or entities are "pure" if they are substantially free of other components. For example, formulations comprising more than about 90% of a particular substance and/or entity are generally considered pure. In some embodiments, substances and/or entities are at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% pure.

Shear force: As used herein, “shear force” refers to a force parallel to the surface of a material as opposed to a force normal to the surface of the material. In some embodiments, the composition has been exposed to high shear forces to create a homogeneous nanoparticle composition. High shear forces can be made using any method known in the art. In some embodiments, cavitation is used to create high shear forces. In some embodiments, high-pressure homogenization is used to create high shear forces. Alternatively or additionally, high shear forces may be applied by exposure to high pressure, for example about 15,000 psi. In some embodiments, such high pressure is in the range of about 18,000 to about 26,000 psi; In some embodiments, the high pressure is in the range of about 20,000 to about 25,000 psi. In some embodiments, a Microfluidizer Processor (Microfluidics Corporation/MFIC Corporation) or other similar device used to produce high shear forces may be used. Microfluidizer Processors accelerate the composition through microchannels (typically having a diameter of 75 microns) at high speeds (typically 50 m/s to 300 m/s) to reduce size to the nanoscale range, resulting in high pressure and resulting high shear rates. Provides. As the fluid exits the microchannel, a jet is formed, which collides with the jet in the opposing microchannel. The fluid in the channel experiences high shear (up to 10 ⁷ l/s), which is much higher than that of the conventional art. It is mixed at the submicron level by jet impingement. Thus, in such an apparatus, it will be possible to obtain particle size reduction and multiphase mixing with high shear and/or impact. In some embodiments of the present invention, the sample is exposed to high shear for a time of less than about 10 minutes. In some embodiments, the time is less than about 9 minutes, about 8 minutes, about 7 minutes, about 6 minutes, about 5 minutes, about 4 minutes, about 3 minutes, about 2 minutes, or about 1 minute. In some embodiments, the time ranges from about 1 to about 2 minutes or less: in some embodiments, the time is about 30 seconds. In some embodiments of the invention, the sample is “microfluidized” through a single exposure to high shear forces; This embodiment is referred to as “one pass” microfluidization.

Small molecule: Generally, a small molecule is understood to mean an organic molecule with a particle size of less than about 5 kd (kilodalton). In some embodiments, the small molecule has a size of less than about 3 Kd, about 2 Kd, or about 1 Kd. In some embodiments, the small molecule has a size of less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, small molecules are non-polymers. In some embodiments, the small molecule is not a protein, peptide, or amino acid. In some embodiments, the small molecule is not a nucleic acid or nucleotide. In some embodiments, the small molecule is not a saccharide or polysaccharide.

Subject: As used herein, “subject” or “patient” refers to any organism to which the composition of the present invention will be administered for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical individuals include animals (e.g., mice, field mice, rabbits, mammals such as non-human primates, humans: insects; worms, etc.).

Substantially: As used herein, “substantially” refers to a quantitative state that represents a degree or magnitude equivalent to all or nearly all of the feature or property of interest. Those of skill in the art will understand that it is extremely rare that biological and chemical phenomena go completely or proceed completely, or that absolute results can be obtained or avoided. “Substantial” is used in many biological and chemical phenomena to capture potential lack of original integrity.

Stable: When used on nanoparticles, “stable” means to maintain one or more aspects of their physical structure (eg, particle size range and/or particle distribution) over time. . In some embodiments of the present invention, stable nanoparticle compositions are those whose average particle size, maximum particle size, particle size range, and/or particle size distribution (e.g., the proportion of particles beyond a specified size) are maintained for a period of time. . In some embodiments, the time is at least about 1 hour; In some embodiments, the time is about 5 hours, about 10 hours, about 1 day, about 1 week, about 2 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months. , May be about 8 months, about 10 months, about 12 months, about 24 months, or even longer. In some embodiments, the time ranges from about 1 day to about 24 months, from about 2 weeks to about 12 months, from about 2 months to about 5 months, and the like. For example, if the nanoparticle composition undergoes long-term storage, temperature change, and/or pH change, if the majority of the nanoparticles in the population remain in diameter within the stated range (e.g., about 10 nm-120 nm), The nanoparticle composition is stable. In this population, most are about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5%. , About 99.6%, about 99.7%, about 99.8%, about 99.9%, or more. In some embodiments of the present invention, when the nanoparticle composition consists of one or more biologically active substances (e.g., unmodified peptides), the concentration of the biologically active substance is maintained in the composition beyond a specified time under specified conditions. , The nanoparticle composition is considered to be stable.

Substantially free of: The nanoparticle composition of the present invention is said to be "substantially free" of particles outside the stated diameter when there are no more than about 50% of the particles outside the stated particle range in the composition. In some embodiments, no more than 25% of the particles are outside this range. In some embodiments, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, Particles of 4%, 3%, 2%, 1%, 0.5% or less have a diameter outside that range.

Suffering from: An individual who is “suffering” with a disease, disease or condition (eg, facial wrinkles) is diagnosed with or has signs of the disease, disease or condition.

Therapeutically effective amount: As used herein, “therapeutically effective amount” is administered to an individual suffering or capable of suffering from a disease, disease, and/or condition. When it refers to a nanoparticle composition in an amount sufficient to treat a disease, disease and/or condition.

Therapeutic Substance: As used, “therapeutic substance” refers to any substance that, when administered to a subject, induces a desired biological and/or pharmacological effect or has a therapeutic effect.

Treatment: As used, “treatment” refers to a partial or complete alleviation, reduction, alleviation, inhibition, delay or degree of onset of one or more features or signs of a particular disease, disease and/or condition. Reduction, refers to any administration of a biologically active ingredient that reduces the incidence. Such treatment can be made in individuals without signs of the associated disease, disease and/or condition, or in individuals exhibiting only initial signs of such disease, disease and/or condition. Alternatively or additionally, such treatment may be in an individual exhibiting one or more established signs of an associated disease, disease and/or condition.

Toxic solvent: As used herein, “toxic solvent” refers to any substance that can alter, rupture, remove or destroy animal tissue. It will be appreciated by those skilled in the art that animal tissue includes living cells, dead cells, extracellular matrix, cell junctions, biological molecules, and the like. Toxic solvents include dimethyl sulfoxide, dimethyl acetate, dimethyl foramide, chloroform, tetramethyl foramide, acetone, acetate, and alkanes, to name a few.

Uniform: The term “homogeneous” as used herein when referring to a nanoparticle composition means that the individual nanoparticles have a specific specified size distribution. For example, in some embodiments, the homogeneous nanoparticle composition has a difference between the minimum and maximum diameters of approximately 600 nm, approximately 550 nm, approximately 500 nm, approximately 450 nm, approximately 400 nm, approximately 350 nm, approximately 300 nm, approximately 250 nm, approximately 200 nm. , Does not exceed about 150 nm, about 100 nm, about 90 nm, about 80 nm, about 70 nm, about 60 nm, about 50 nm, or less than that. In some embodiments, the particles (e.g., particles comprising an unmodified-peptide) in the nanoparticle composition of the present invention are about 600 nm, about 550 nm, about 500 nm, about 450 nm, about 400 nm, about 350 nm, about 300 nm, It has a diameter of about 250 nm, about 200 nm, about 150 nm, about 130 nm, about 120 nm, about 115 nm, about 110 nm, about 100 nm, about 90 nm, about 80 nm or less. In some embodiments, particles (eg, particles comprising an unmodified-peptide) in nanoparticle compositions of the present invention have a diameter in the range of about 10 nm to about 600 nm. In some embodiments, particles (e.g., particles comprising an unmodified-peptide) in the nanoparticle composition of the present invention are from about 10 nm to about 300 nm, from about 10 nm to about 200 nm, from about 10 to about 150 nm, from about 10 nm to about 10 nm. It has a diameter in the range of about 130 nm, about 10 nm to about 120 nm, about 10 nm to about 115 nm, about 10 nm to about 110 nm, about 10 nm to about 100 nm, or about 10 nm to about 90 nm. In some embodiments, particles (e.g., particles comprising an unmodified-peptide) in the nanoparticle composition of the present invention are about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 130 nm, about 120 nm, about 115 nm, It has an average particle size of less than about 110 nm, about 100 nm, or about 90 nm. In some embodiments, the average particle size ranges from about 10 nm to about 300 nm, about 50 nm to about 250 nm, about 60 nm to about 200 nm, about 65 nm to about 150 nm, about 70 nm to about 130 nm. In some embodiments the average particle size is about 80 nm to about 110 nm. In some embodiments the average particle size is about 90 nm to about 100 nm. In some embodiments, the majority of particles (eg, particles comprising an unmodified-peptide) within the homogeneous nanoparticle composition of the present invention are within a specified size or within a specified range. In some embodiments, the majority of the particles of the composition are 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or more. In some embodiments of the present invention, a homogeneous nanoparticle composition is obtained by microfluidization of a sample. In some embodiments of the present invention, homogeneous nanoparticle compositions can be prepared by exposure to high shear forces such as microfluidization.

Unmodified Peptide: As used herein, “unmodified peptide” refers to a peptide that is not chemically modified by adding another covalently bonded functional group with the intention of transdermal transport of the peptide. In some embodiments, the peptide has not been chemically modified by adding pendant acetyl and/or palmitoyl groups. In some embodiments, the peptide has not been chemically modified by adding an optional functional pendant group.

Figure 1. Histological analysis of mice treated with peptide nanoparticles. Photomicrographs of skin tissues colored with Masson's Trichrome pigmentation are shown. The average histological score is 2.33 of 4 total for the control (nanoparticles prepared without pentapeptide). The average histological score of the treatment group (prepared nanoparticles with pentapeptide) was 3.67 of the total 4.

Description of specific embodiments

Nanoparticles

As described herein, the present invention provides nanoparticle compositions comprising one or more unmodified peptides. In some embodiments, such nanoparticle compositions further comprise one or more biologically active substances in addition to the unmodified peptide. In some embodiments, nanoparticle compositions are formulated with one or more additional ingredients, such as pharmaceutical or cosmetic compositions. In some embodiments, such pharmaceutical or cosmetic compositions are formulated to allow transdermal delivery of unmodified peptides (and/or one or more biologically active substances).

In some embodiments, the nanoparticle compositions of the present invention are stable. In some embodiments, the nanoparticle composition is homogeneous. The difference between the minimum and maximum diameters does not exceed about 600 nm, about 550 nm, about 500 nm, about 450 nm, about 400 nm, about 350 nm, about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 100 nm.

In some embodiments, the nanoparticles of the present invention are about 1000 nm, about 600 nm, about 550 nm, about 500 nm, about 450 nm, about 400 nm, about 350 nm, about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 130 nm, about 120 nm, It has a diameter of about 115 nm, about 110 nm, about 100 nm, about 90 nm, about 80 nm, about 50 nm, or less.

In some embodiments, the nanoparticles of the present invention are 1 nm to 1000 nm, 1 nm to 600 nm, 1 nm to 500 nm, 1 nm to 400 nm, 1 nm to 300 nm, 1 nm to 200 nm, 1 nm to 150 nm, It has a diameter of 1 nm to 120 nm, 1 nm to 100 nm, 1 nm to 75 nm, 1 nm to 50 nm, or 1 nm to 25 nm. In some embodiments, the nanoparticles of the present invention have a diameter of 1 nm to 15 nm, 15 nm to 200 nm, 25 nm to 200 nm, 50 nm to 200 nm, or 75 nm to 200 nm.

In some embodiments, the overall particle distribution is within a specific diameter size range. In some embodiments, less than 50%, 25%, 10%, 5%, or 1% of the total particle distribution is outside the specified specific particle size distribution. In some embodiments, the nanoparticle composition is substantially free of particles with a diameter greater than 300 nm, 250 nm, 200 nm, 150 nm, 120 nm, 100 nm, 75 nm, 50 nm, or 25 nm.

In some embodiments, the nanoparticle composition has an average particle size of about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 130 nm, about 120 nm, about 115 nm, about 110 nm, about 100 nm, about Less than 90 nm, or about 50 nm. In some embodiments, the average particle size ranges from about 10 nm to about 300 nm, 50 nm to about 250 nm, 60 nm to about 200 nm, 65 nm to about 150 nm, or 70 nm to about 130 nm. In some embodiments, the average particle size is about 80 nm to about 110 nm. In some embodiments, the average particle size is about 90 to about 100 nm.

In some embodiments, nanoparticle compositions of the present invention are substantially free of particles with a diameter greater than 300 nm. In particular, in some embodiments, less than 50% of the nanoparticles of the nanoparticle composition of the present invention exceed 300 nm. In some embodiments, less than 25% of the nanoparticles of the nanoparticle composition of the present invention exceed 300 nm. In some embodiments, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, nanoparticles of the nanoparticle composition of the present invention, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5% or less exceed 300nm.

Additionally, in some embodiments, the nanoparticles in the nanoparticle compositions of the present invention have a diameter in the range of 10 nm to 300 nm.

In some embodiments, nanoparticle compositions of the present invention are substantially free of particles greater than 200 nm. In particular, in some embodiments, less than 50% of the nanoparticles in the nanoparticle composition of the present invention exceed 200 nm in diameter. In some embodiments, less than 25% of the nanoparticles in the nanoparticle composition of the present invention exceed 200 nm in diameter. In some embodiments, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5 of the particles %, 4%, 3%, 2%, 1%, less than 0.5% or less have a diameter in excess of 200 nm. Further, in some embodiments, the nanoparticles in the nanoparticle compositions of the present invention have a diameter in the range of 10 nm to 200 nm.

In some embodiments, nanoparticle compositions of the present invention are substantially free of particles greater than 120 nm. In particular, in some embodiments, less than 50% of the nanoparticles in the nanoparticle composition of the present invention exceed 120 nm in diameter. In some embodiments, less than 25% of the nanoparticles in the nanoparticle composition of the present invention exceed 120 nm in diameter. In some embodiments, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5 of the particles %, 4%, 3%, 2%, 1%, less than 0.5% or less have a diameter in excess of 120 nm. Further, in some embodiments, the nanoparticles in the nanoparticle compositions of the present invention have a diameter in the range of 10 nm to 200 nm.

In some embodiments, the majority of nanoparticles in the composition of the present invention have a diameter of a specific size or less or a specified range. In some embodiments, most of the particles in the composition are 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, Refers to 99.6%, 99.7%, 99.8%, 99.9% or more.

Zeta potential is a measure of the electric potential in the shear plane. The shear plane is an imaginary plane that separates a thin layer of liquid bound to a solid plane (for example, the nanoparticle surface of the present invention), and exhibits elastic behavior from the remaining liquid (for example, a liquid dispersion medium) exhibiting normal viscous behavior. In some embodiments, nanoparticles of the invention have a zeta potential ranging from -50 mV to +50 mV. In some embodiments, nanoparticles of the invention have a zeta potential in the range of -25 mV to +25 mV. In some embodiments, nanoparticles of the invention have a zeta potential in the range of -10 mV to +10 mV.

In some embodiments, the nanoparticle composition of the present invention is an emulsion or dispersion. In general, a dispersion or emulsion is formed by a combination of at least two unmixed substances, one of which will constitute a medium in which particles (eg, nanoparticles, constituting a “dispersed medium”) are dispersed. Hydrophilic emulsified ("oil-in-water") dispersions are those in which oily particles (or hydrophobic or non-polar) are dispersed in an aqueous dispersion medium. Lipophilic emulsification ("water-in-oil") is the dispersion of water-soluble (or hydrophilic or polar) particles in an oily dispersion medium. Those skilled in the art will understand that dispersions can be formed from any two immiscible media, but are not strictly limited to water-soluble and oily media combinations. The “dispersion medium” is widely applied as any dispersion medium, although it is usually referred to in the “aqueous” and “oily” categories, for example, emulsions or dispersions can be mixed. No hydrophobic/hydrophilic; can be prepared from polar/non-polar materials, irrespective of what such materials are strictly distinguished as'water-soluble' or'oil'.

In some embodiments, nanoparticle compositions of the present invention comprise micelle structures (the nanoparticles are micelles). In some embodiments, such micelle structures are cross-linked. In some embodiments, such micelle structures are not cross-linked.

In some embodiments, nanoparticle compositions of the present invention are self-assembled from a complex collection of components. In some embodiments, nanoparticle compositions of the present invention can be prepared by exposing the component complex (eg “premix”) to high shear forces. In some embodiments, high shear forces are provided by high pressure, cavitation, homogenization and/or microfluidization. In some embodiments, the combined nanoparticle-forming component is perturbed, agitated, or otherwise mixed. In some such embodiments, the ingredients are subjected to high shear forces after mixing. In some specific embodiments, mixing can be performed for less than 1 hour or for more than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 hours. In some embodiments, solubilization occurs.

In some embodiments of the present invention, production of nanoparticle compositions involves dialyzing and freeze-drying a collection of ingredients to remove any organic solvent to make the composition.

It will be appreciated that in some embodiments of the present invention utilizing a premix, the premix component can be assembled prior to providing a high shear force. At least some of these particles may be microparticles or even nanoparticles. In some embodiments, the nanoparticle composition of the present invention is prepared from a premix, the premix being selected from a suspension or a microecho. However, in some embodiments, the particulate structures do not form a premix before providing a high shear force.

In some embodiments of the present invention, all components present in the final nanoparticle composition are present in the premix, and these components are subjected to high shear forces to form the nanoparticle composition. In some embodiments of the invention, one or more components present in the final nanoparticle composition are either left out of the premix or present in the premix in an amount less than the amount present in the final nanoparticle composition. That is, in some embodiments, one or more substances are added to the nanoparticle composition after the premix is subjected to high shear forces.

In certain embodiments of the present invention, the premix is prepared as a solution prior to providing a high shear force. In particular, in the case of nanoparticle compositions comprising at least one biologically active substance (eg, an unmodified peptide), it is sometimes desirable to dissolve the biologically active substance in the premix before high shear forces are provided. Thus, in many embodiments, the biologically active substance is soluble in at least one medium (or complex medium used in the premix). In some embodiments, heat is required for such dissolution, but not in other embodiments.

In some embodiments of the invention, the nanoparticle composition comprises one or more water-soluble, polar or hydrophilic media, one or more oily, non-polar, or hydrophobic media, one or more micelle components, one or more surfactants, or It is made from ingredients containing emulsifiers, one or more biologically active substances and/or one or more delaying release substances.

Those skilled in the art will be well aware of suitable aqueous media that can be used as dispersion media or media to be dispersed according to the invention. Representative aqueous media include, for example, water, salt solution (phosphate buffer salt), water for injection, short chain alcohol, 5% dextrose, Ringer's solution (lactate Ringer's injection, lactate Ringer's injection + 5% dextrose, ah. Ringer's injection), Normosol_M, Risolite E and the like, and combinations thereof.

Those skilled in the art will be well aware of suitable oily media that can be used as dispersion media or media to be dispersed according to the invention. In some embodiments, the oil may be composed of one or more fatty acid groups or salts thereof. In some embodiments, fatty acid groups may be composed of digestible, long chain (eg, C ₈ -C ₅₀ ), substituted or unsubstituted hydrocarbons. In some embodiments, the fatty acid group _{can be a C 10} -C ₂₀ fatty acid or salt thereof. In some embodiments, the fatty acid group _{can be a C 15} -C ₂₀ fatty acid or salt thereof. In some embodiments, the fatty acid group _{can be a C 15} -C ₂₅ fatty acid or salt thereof. In some embodiments, the fatty acid group may be unsaturated. In some embodiments, the fatty acid group may be a monounsaturated acid. In some embodiments, the double bond of the unsaturated fatty acid group may be in the cis form. In some embodiments, the double bond of the unsaturated fatty acid group may be in the trans form.

In some embodiments, the fatty acid group may be one or more of butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palytic acid, stearic acid, arachidic acid, behenic acid, or rhonoseric acid. have. In some embodiments, the fatty acid group is one or more palmitoleic acid, oleic acid, vaccenic acid, linoleic acid, alpha-linolenic acid. linolenic acid), gamma-linoleic acid, arachidonic acid, gadoleic acid, arachidonic acid, eicosapentaenoic acid, doco It may be docosahexaenoic acid or erucic acid.

In some embodiments, the oil is a liquid triglyceride. In some embodiments, the oil is a medium chain (e.g., 6-12 carbon) triglyceride (e.g. Labrafac WL 1349, coconut oil, palm kernel oil, camphor tree drupe oil), Etc). In certain embodiments, the oil is a short chain (eg 2-5 carbon) triglyceride. In certain embodiments, the oil is a long chain (eg 12 or more carbons) triglycerides (eg, soybean oil, sunflower oil, etc.).

Suitable oils that can be used in the present invention include almonds, apricot kernels, avocados, babassu palm, bergamot, black current seeds, borage, cade, chamomile, canola. ), caraway, Brazilian carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cottonseed, Emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop , Isopropyl myristate, jojoba, kukui nut, labandin, lavender, lemon, litsea cubeba, macademia nut, mallow, Mango seeds, meadowfoam seeds, minerals, mink, nutmeg, olives, oranges, orange roughy, palms, palm kernels, peach kernels, peanuts , Poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savory, mountain birch sea buckthorn), sesame seeds, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, malt and mixtures thereof. Does not. Suitable synthetic oils that can be used in the present invention include caprylic/capric triglycerides, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, octyldodecanol, oleyl alcohol and combinations thereof. It is not limited to this.

Suitable micelle components may include, for example, one or more amphiphilic entities. Useful amphoteric entities include natural entities, synthetic entities, and entities containing both natural and synthetic components. In some embodiments, the amphoteric entity may be composed of one or more polymers, and/or one or more compounds having polymeric properties.

In general, amphoteric entities are those that have both hydrophobic and hydrophilic properties. One of skill in the art will recognize that an amphoteric entity can be composed of a number of different things. In some embodiments, an amphoteric entity may be composed of one or more individual compounds or molecules that are amphoteric themselves. For example, such compounds or molecules include polyethylene glycol (PEG), phospholipids, cholesterol, glycolipid fatty acids, bile acids, and saponins. PEG is generally approved by the US FDA as safe for use in food, cosmetics and pharmaceuticals. PEG is water soluble, non-toxic, colorless, lubricious, non-volatile and non-irritating.

In some embodiments, an amphoteric entity is one or more individual components that are not amphoteric by themselves but have some hydrophilic or hydrophobic properties. In such an embodiment, two or more such non-amphipathic components are generally associated with others so that the population of individual components is amphoteric. Covalent bonds may or may not be involved in such associations, and such associations may or may not be related to non-covalent bonding (e.g., electrical interactions, affinity interactions, hydrophobic interactions, hydrogen bonding, van der Waals interactions, ionic interactions). Action, dipole-dipole interaction, etc.) may be involved. In general, any associated force, bonding or adsorption means may be involved in such association.

In some embodiments, the amphoteric entity according to the present invention may be made of two or more individual components with different hydrophobicity or hydrophilicity. In certain embodiments, the amphoteric entity may consist of at least one hydrophilic component and at least one hydrophobic component. In certain embodiments, the “hydrophilic” and “hydrophobic” components are either hydrophilic or hydrophobic with respect to each other.

In some embodiments, two or more components having different degrees of hydrophilicity or hydrophobicity may be covalently bonded to form a homopolymer or co-polymer. In some embodiments, the co-polymer may be a block co-polymer. In some embodiments, the co-polymer may be a graft co-polymer.

In some embodiments, the amphoteric entity may be composed of an amphoteric block co-polymer. In some embodiments, the amphoteric block co-polymer may be a diblock co-polymer. In certain embodiments, in the amphoteric diblock co-polymer, the first polymer block and the second polymer block are covalently bonded to each other at the ends of the chain. In certain embodiments, the first polymer block may be composed of repeating units of a hydrophilic component, and the second polymer block may be composed of repeating units of a hydrophobic component. In certain embodiments, the first polymer block may be composed of repeating units of a hydrophobic component, and the second polymer block may be composed of repeating units of a hydrophilic component. In some embodiments, the amphoteric block co-polymer may be a multiblock co-polymer.

In certain embodiments, the amphoteric block co-polymer may consist of multiple blocks in which alternating blocks of two or more polymers are covalently linked at the ends of a chain. In certain embodiments, the amphoteric block co-polymer may be composed of multiple blocks of alternating hydrophilic blocks and hydrophobic blocks covalently linked at the ends of the chain. In certain embodiments, each alternating block may be composed of a hydrophilic component or a repeating unit of hydrophobic components.

In some embodiments, the amphoteric entity may comprise or consist of an amphoteric graft co-polymer. In some embodiments, the amphoteric graft co-polymer may comprise or be configured such that a block of the polymer is covalently linked to the side chain of another block of the polymer. In certain embodiments, each polymer block may be comprised or comprised of repeat units of hydrophilic or hydrophobic components. In certain embodiments, the amphoteric graft co-polymer may comprise or consist of a first polymer block and a polymer block covalently linked to the side chains of the first polymer block. In certain embodiments, the first polymer block may include or consist of repeat units of a hydrophilic component, and the second block may include repeat units of a hydrophobic component. In certain embodiments, the first polymer block may include a repeating unit of a hydrophobic component, and the second block may include a repeating unit of a hydrophilic component.

In some embodiments, the amphoteric block or graft co-polymer may include a hydrophilic polymer block comprising repeating units of polysaccharide and a hydrophobic polymer block comprising repeating units of polyester or polysaccharide.

Alternatively or additionally, the amphoteric block or graft co-polymer may comprise a hydrophobic polymer block comprising repeating units of polysaccharide and a hydrophilic polymer block comprising repeating units of polyester or polysaccharide. Such hydrophilic polymer blocks include repeating units of any type of hydrophilic polymer, such as polysaccharides (e.g., pullulan) or polyalkene oxides (e.g., polyethylene oxide). May be. The hydrophobic polymer block may contain a hydrophobic polymer repeating unit such as polycaprolactone or polyamide (eg, polycaplactam).

In some embodiments, the hydrophilic portion of the amphoteric entity may be non-ionic. In some embodiments, the hydrophilic portion of the amphoteric entity may be one or more ionic groups. In general, such ionic groups are hydrophilic and can impart hydrophilicity to amphoteric entities.

In some embodiments, the ionic group may be a cationic group. In some embodiments, the cationic group is ammonium (NH ₄ ⁺ ), nitronium (NO ₂ ⁺ ), ^{nitrosyl (NO +} ), hydronium (H ₃ O ⁺ ), first mercury (Hg ₂ ^{2 +} ) , Phosphonium (PH ₄ ⁺ ), vanadyl (VO ^{2 +} ), or may be a salt thereof.

In some embodiments, the ionic group may be an anionic group. In some embodiments, the anionic group is a fatty acid, arsenic (As ^{3 -),} azide (N ₃ ^-), bromine-water (Br ^-), chloride (Cl ^-), fluoride (F ^-), hydride (H ^-), iodide (I ^-), nitride (N ^{3 -),} oxide (O ^{2 -),} phosphide (P ^{3 -),} celecoxib Need (Se ^2-), sulfide (S ^{2 -),} a peroxide (O _{² 2} ^-) , arsenate (AsO ₄ ^{3 -),} arsenite (AsO _{³ 3} ^-), borate (BO ₃ ^3-), and bromate (BrO ₄ ^-), bromates (BrO ₃ ^-), O cancel acid (BrO ₂ ^-) , hypochlorous canceled acid (BrO ^-), carbonate (CO ₃ ^{2 -),} bicarbonate (HCO ₃ ^-), chlorate (ClO ₃ ^-), perchlorate (ClO ₄ ^-), chlorate (ClO ₂ ^-), hypochlorite (ClO ^-), chromate (CrO ₄ ^{2 -),} dichromate _{^{(Cr 2 O 7 2 -)}} , perfluoroalkyl acid salt (BrO ₄ ^-), fluoride salts (BrO ₃ ^-), O fluoride salt (BrO ₂ ^-) , hypochlorous fluoride salt (BrO ^-), periodate salts (IO ₄ ^-), iodide salts (IO ₃ ^-), O iodine salt (IO ₂ ^-), hypochlorous iodine salt (IO ^-), nitrate (NO ₃ ^-), nitrite (NO ₂ ^-), phosphate (PO ₄ ^{3 -),} phosphate Flame (HPO ₄ ^{2 -),} phosphate Flame _{^{(H 2 PO 4 2 -)}} , phosphites (PO _{³ 3} ^-), silicate (SiO ₃ ^{2 -),} sulfate (SO ₄ ^2-), thiosulfate (S ₂ O ₃ ^2-), hydrogen sulfide salts (HSO ₄ ^-), sulfite (SO ₃ ^2-), sulfite flame (HSO ₃ ^{2 -),} sulfonate _{(-S (= O) 2 -0} -), acetate ^{_{(C 2 H 3 O 2 -}} ), formic acid (HCO ₂ ^-), oxalate _{^{(C 2 O 4 2 -)}} , hydrogen oxalate (HC ₂ O ₄ ^-), citrate (C6H5O7 3), succinic acid ^{_{(C 4 H 4 O 4 2}} -), fumarate ^{_{(C 4 H 2 O 4 2}} -), maleate ^{_{(C 4 H 5 O 5 2}} -), hydrogen sulfide (HS ^-), telru Le (Te ^{2 -),} amide (NH ^{2 -),} cyanate (OCN ^-), thiocyanate (SCN ^-), cyanide (CN ^-), hydroxide (OH ^-), permanganate (MnO ₄ ^-), Or it may be a salt thereof.

In some embodiments, the hydrophilic component of the amphoteric entity comprises or consists of a nucleic acid. For example, the nucleic acid polymer may include DNA, RNA, or a combination thereof. In some embodiments, the nucleic acid polymer may be an oligonucleotide and/or a polynucleotide. In some embodiments, the nucleic acid polymer comprises oligonucleotides and/or modified oligonucleotides; Antisense oligonucleotides and/or modified antisense oligonucleotides; cDNA; Genomic DNA; Viral DNA and/or RNA; DNA and/or RNA chimera; Plasmid; Cosmid; Gene fragments; Artificial and/or natural chromosomes (eg, yeast artificial chromosomes) and/or portions thereof; RNA (eg, mRNA, tRNA, rRNA and/or ribozyme); Peptide nucleic acid (PNA); Polynucleotides composed of modified or unmodified synthetic nucleic acid analogs; A variety of DNA structures including single false DNA, double stranded DNA, supercoiled DNA and/or triple-helix DNA; Z-DNA; And/or a combination thereof.

In some embodiments, the hydrophilic component of the amphoteric entity consists of or comprises a carbohydrate. In some embodiments, the carbohydrate is a polysaccharide to which simple sugars (or derivatives thereof) are linked by glycosidic bonds, as known in the art. Such sugars include glucose, fructose, galactose, ribose, lactose, sucrose, maltose, trehalose, celbiose, mannose, silose, arabinose, glucuronic acid, galactoronic acid, mannuronic acid. , Glucosamine, galactosamine, neuramic acid, etc. are included, but are not limited thereto. In some embodiments, the polymer can be a hydrophilic carbohydrate including aminated, carboxylated and sulfated polysaccharides. In some embodiments, the hydrophilic carbohydrate is one or more of pullulan, cellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, hydroxycellulose, methylcellulose, dextran, cyclodextran, glycogen, starch, hydroxyethyl starch, Carrageenan, glycol, amylose, chitosan, N,O-carboxylmethylchitosan, algin and alginic acid, starch, chitin, heparin, konzak, glucomannan, putturan, hepamine, hyaluronic acid, kurdran and xanthan, etc. Can be In some embodiments, the hydrophilic polysaccharide may be modified to be hydrophobic by introducing a large number of side chain hydrophobic groups. In some embodiments, the hydrophobic carbohydrate may also include cellulose acetate, pullulan acetate, Konzac acetate, amylose acetate, dextran acetate, and the like.

In some embodiments, the hydrophilic component of amphoteric entini includes, but is not limited to, xanthan gum, alginic acid, karaya gum, alginate sodium and/or locust bean gum, and the like.

In some embodiments, a component of an amphoteric entity may comprise or consist of a protein. In some embodiments, the protein is a hydrophilic component of an amphoteric entity. In another embodiment, the protein is a hydrophobic component of an amphoteric entity. Exemplary proteins that can be used in the present invention include, but are not limited to, albumin, collagen, or poly(amino acids) (eg, polylysine).

In some embodiments, the hydrophobic component of the amphoteric entity may comprise or consist of one or more fatty acid groups or salts thereof. In general, such groups can impart hydrophobic properties on amphoteric entities. In some embodiments, fatty acid groups may be composed of long degradable chains (eg, C ₈ -C ₅₀ ), substituted or unsubstituted hydrocarbons. In some embodiments, the fatty acid group _{can be a C 10} -C ₂₀ fatty acid or salt thereof. In some embodiments, the fatty acid group _{can be a C 15} -C ₂₀ fatty acid or salt thereof. In some embodiments, the fatty acid group _{can be a C 15} -C ₂₅ fatty acid or salt thereof. In some embodiments, the fatty acid group may be unsaturated. In some embodiments, the fatty acid group may be a monounsaturated acid. In some embodiments, the double bond of the unsaturated fatty acid group may be in the cis form. In some embodiments, the double bond of the unsaturated fatty acid group may be in the trans form.

In some embodiments, the fatty acid group may be one or more of butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palytic acid, stearic acid, arachidic acid, behenic acid, or rhonoseric acid. have. In some embodiments, the fatty acid group is one or more of palmitoleic acid, oleic acid, basenic acid, linoleic acid, alpha-linolenic acid, gamma-linoleic acid, arachidonic acid, gadoleic acid, arachidonic acid, eico It may be sapentaenoic acid, docosahexaenoic acid or erucic acid.

In some embodiments, the hydrophobic component of the amphoteric entity is one or more biocompatible and/or biodegradable synthetic polymers such as polycarbonate (e.g., poly(l,3-dioxane-2one) ), polyanhydride (e.g., poly(seba-type anhydride)), polyhydroxy acid (e.g., poly(-hydroxyalkanoate)), polypropyl fumarate, polycarporactone , Polyamides (e.g. polycarrolactam), polyacetals, polyethers, polyesters (e.g. polylactide and polyglycolide), biodegradable polysaanoacrylate, polyvinyl alcohol and biodegradable It can be polyurethane or the like. For example, the amphoteric entity may comprise one or more of the following biodegradable polymers: poly(lactic acid), poly(glycolic acid), poly(caprolactone), poly(lactide-co-glycolide) , Poly(lactide-co-caprolactone), poly(glycolide-co-caprolactone), and poly(DL-lactide-co-glycolide).

In some embodiments, the hydrophobic component of the amphoteric entity may comprise or consist of one or more acrylic polymers. In certain embodiments, acrylic polymers include, for example, acrylic and methacrylic acid co-polymers, methyl methacrylate co-polymers, ethoxyethyl methacrylate, cyanoethyl methacrylate, aminoalkyl methacrylate co- Polymer, poly(acrylic acid), poly(methacrylic acid), methacrylic acid alkylamide co-polymer, poly(methyl methacrylate), poly(methacrylic acid anhydride), methyl methacrylate, polymethacrylate, Poly(methyl methacrylate) co-polymer, polyacrylamide, aminoalkyl methacrylate co-polymer, glycidyl methacrylate co-polymer, and a composite comprising one or more of the aforementioned polymers may be included. have. The acrylic polymer may include fully-polymerized acrylic and methacrylic acid ester co-polymers having a low content of tetravalent ammonium groups.

In some embodiments, the hydrophobic component of the amphoteric entity may comprise or consist of polyester. Examples of such polyesters include polyalkylene glycol, poly(glycolide-co-lactide), PEGylated poly(latic-co-glycolic acid), poly(lactic acid), PEGylated Copolymers of poly(lactic acid), poly(glycolic acid), PEGylated poly(glycolic acid), polylactic and polyglycolic acid, and derivatives thereof may be included. In some embodiments, polyesters include polyanhydride, poly(ortho ester) PEGylated poly(ortho ester), poly(caprolactone), pegylated poly(caprolactone), polylysine, Pegylated polylysine, poly(ethylene imine), pegylated poly(ethylene imine), and derivatives thereof may also be included. In some embodiments, polyesters include, for example, polycaprolactone, poly(L-lactide-co-L-lysine), poly(serine ester), poly(4-hydroxy-L-proline ester), poly [α-(4-aminobutyl)-L-glycolic acid] and derivatives thereof may also be included.

Suitable surfactants or emulsifiers include phosphoglycerides; Phosphatidylcholine; Dipalmitoyl phosphatidylcholine (DPPC); Dioleyylphosphatidyl ethanolamine (DOPE); Dioleyloxypropyltriethylammonium (DOTMA); Dioleylphosphatidylcholine; cholesterol; Cholesterol esters; Diacylglycerol; Diacylglycerol succinate; Diphosphatidyl glycerol (DPPG); Hexanedecanol; Polyethylene glycol (PEG); Fatty acids such as polyoxyethylene-9-lauryl ether; Surface active fatty acids such as palmitic acid or oleic acid; fatty acid; Fatty acid amide; Sorbitan trioleate (Span 85) glycocholate; Sorbitan monolaurate (Span 20); Polysorbate 20 (Tween-20); Polysorbate 60 (Tween-60); Polysorbate 65 (Tween-65); Polysorbate 80 (Tween-80); Polysorbate 85 (Tween-85); Polyoxyethylene monostearate; Sulpectin; Poloxomer; Sorbitan fatty acid esters such as sorbitan trioleate; lecithin; Lysolecithin; Phosphatidylserine; Phosphatidylinositol; Sphingomyelin; Phosphatidylethanolamine (cephaline); Cardiolipin; Phosphatidic acid; Cerebroside; Dicetyl phosphate; Dipalmitoylphosphatidylglycerol; Stearylamine; Dodecylamine; Hexadecyl-amine; Acetyl palmitate; Glycerol ricinoleate; Hexadecyl stearate; Tiloxapol; Poly(ethylene glycol)5000-phosphatidylethanolamine; Poly(ethylene glycol)400-monostearate; And phospholipids, but are not limited thereto.

The surfactant component may be a mixture of different surfactants. These surfactants can be extracted and purified from natural sources, or they can be synthesized in the laboratory. In a suitable embodiment, a commercially available surfactant may be used.

In certain embodiments of the present invention, nanoparticles having desired properties can be made by selecting and adjusting the relative amounts of ingredients used to prepare the nanoparticle compositions of the present invention. In some embodiments, the oil and surfactant are used in a ratio ranging from 0.25-10. In some embodiments, the ratio of oil to surfactant is approximately 0.25:1, approximately 0.5:1, approximately 1:1, approximately 2:1, approximately 3:1, approximately 4:1, approximately 5:1, approximately 6 :1, about 7:1, about 8:1, about 9:1, or about 10:1. In some embodiments, the ratio of surfactant to oil is approximately 0.5:1, approximately 1:1, approximately 2:1, approximately 3:1, approximately 4:1, approximately 5:1, approximately 6:1, approximately 7 :1, about 8:1, about 9:1, or about 10:1. In some embodiments, the ratio of oil to surfactant ranges from 0.25-2. In some embodiments, the ratio of oil to surfactant is approximately 0.25:1, approximately 0.5:1, approximately 1:1, or approximately 2:1. In some embodiments, the ratio of surfactant to oil is approximately 0.5:1, approximately 1:1, or approximately 2:1. In certain embodiments, the ratio of oil to surfactant is approximately 1:1.

In some embodiments, the proportion of oil in the composition (eg, premix) from which the nanoparticles are prepared is between 0% and 30%. The proportion of oil in the composition from which the nanoparticles are prepared (e.g., premix) is approximately 1%, approximately 2%, approximately 3%, approximately 4%, approximately 5%, approximately 6%, approximately 7%, approximately 9%. , About 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, or about 30%. In some embodiments, the proportion of oil is approximately 8%. In some embodiments, the proportion of oil is approximately 5%.

In some embodiments, the proportion of oil in the composition (e.g., premix) from which the nanoparticles are prepared when one or more amphoteric entities are used is 40% to 99%, 50% to 99%, 60% To 99%, 70% to 99%, 80% to 99%, 80% to 90%, or 90% to 99%. In some embodiments, the proportion of amphoteric entities in a composition from which nanoparticles are prepared (e.g., premix) is approximately 75%, approximately 76%, approximately 77%, approximately 78%, approximately 79%, approximately 80%, About 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93 %, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%.

The proportion of the substance having surfactant activity in the premix is 0% to 99%, 10% to 99%, 25% to 99%, 50% to 99%, or 75% to 99%. In some embodiments, the proportion of material having surfactant activity in the premix is 0% to 75%, 0% to 50%, 0% to 25%, or 0% to 10%. In some embodiments, the proportion of surfactant in the premix is 0%-30%. In some embodiments, the proportion of surfactant in the composition is approximately 1%, approximately 2%, approximately 3%, approximately 4%, approximately 5%, approximately 6%, approximately 7%, approximately 9%, approximately 10%, approximately 11 %, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, About 24%, about 25%, about 26%, about 27%, about 28%, about 29%, or about 30%. In some embodiments, the proportion of surfactant is approximately 8%. In some embodiments the proportion of surfactant is approximately 5%.

In some embodiments, the nanoparticle composition does not include one or more oils. In some embodiments, nanoparticle compositions may include two or more oils. In some embodiments, the nanoparticle composition does not include one or more surfactants. In some embodiments, nanoparticle compositions may include two or more surfactants. In some embodiments, the nanoparticle composition is completely or substantially free of toxic components.

In some embodiments, the nanoparticle composition consists essentially of water, oil, surfactant, and at least one biologically active substance (eg, unmodified peptide). In some embodiments, the nanoparticle composition consists essentially of water, an oil, a surfactant, and at least one biologically active material, and at least one material used to produce and/or preserve the nanoparticle composition.

In some embodiments, the nanoparticle composition consists essentially of water, an oil, a surfactant, and an unmodified peptide. In some embodiments, the nanoparticle composition consists of water, an oil, a surfactant, an unmodified peptide, and at least one material used to produce and/or preserve the nanoparticle composition.

Untransformed Peptide

Various peptides can be included in the nanoparticle composition according to the present invention. In most embodiments, the peptide is less than about 100 amino acids in length; In some embodiments, the peptide is about 90 amino acids, about 80 amino acids, about 70 amino acids, about 65 amino acids, about 60 amino acids, about 55 amino acids, about 50 amino acids, about 45 amino acids, about 40 amino acids. Amino acids, about 35 amino acids, about 30 amino acids, about 25 amino acids, about 20 amino acids, about 15 amino acids, about 13 amino acids, about 12 amino acids, about 10 amino acids, about 9 amino acids, about 8 Amino acids, about 7 amino acids, about 6 amino acids, or less than about 5 amino acids. In some embodiments, the peptide is a pentapeptide. In some embodiments, the peptide bound to the nanoparticle composition consists entirely of naturally occurring amino acids. In some embodiments, the peptide consists of one or more naturally occurring amino acids.

Short unmodified peptides used according to the present invention are generally those that have biological activity in the skin (including dermis and epithelium), subcutaneous tissue (including adipose tissue) and/or adjacent muscles. Such peptides include peptides that promote extracellular matrix production (e.g. KTTKS, SEQ ID NO.: 1; EYKTTKSSRL, SEQ ID NO.: 2; VIEYKTTK, SEQ ID NO.: 3; KTTK, SEQ ID NO.: 4; GKTVIEYKTTKS, SEQ ID NO.: 5; GKTVIEYKTTKSSRL, SEQ ID NO.: 6; WGKTVIEYKTTKSSRLPIID, SEQ ID NO.: 7; CTSHTGAWGKTVIEYKTTKS, SEQ ID NO.: 8; TTKS, SEQ ID NO.: 9), wrinkles Peptides that reduce (e.g. EEMQRR, SEQ ID NO.: 10), peptides that improve wound healing (e.g. gastrin-releasing peptide, VGVAPG, SEQ ID NO.: 11; YYRADA, SEQ ID NO.: 12; GHK, SEQ ID NO.: 13, interferon, interferon inducer), and peptides (e.g. For example, P144; TSLDASIIWAMMQN, SEQ ID NO.: 14), including but not limited to (Katayama, et al; supra, Lupo, supra; Robinson et al., supra; Bhartiya et al., 1992, J. Cell). Physiol, 150:312; and Santiago et al, 2005, J. Investigative Dermatology, 125:450; all contents are attached by reference). Refer to Table 1 below for definitions of peptide abbreviations.

Other ingredients

As mentioned herein, the nanoparticle composition of the present invention may contain one or more other components or may be combined. Examples of other ingredients are discussed here.

Biologically active substance

According to the present invention, any biologically active substances may be carried, including therapeutic agents, diagnostic agents, prophylactic agents, nutritional agents, cosmetic agents and/or dermatological substances. Such biologically active substances include small molecules, organometallic compounds, nucleic acids, proteins (multimer proteins, protein complexes, etc.), peptides, lipids, carbohydrates, herbs, hormones, metals, radioactive elements and compounds, drugs, vaccines, immunological It may be a substance and/or a combination thereof. Such biologically active substances may be present in, interspersed, or absorbed or trapped on the surface of the micelle membrane of the nanoparticles of the present invention.

In some embodiments, the proportion of biologically active material in the composition (eg, premix) used to prepare the nanoparticles of the present invention is 0.1%-25%. In some embodiments, the proportion of biologically active material is 0.1%-20%, 0.1%-15%, 0.1%-10%, 0.1% -5%, or 0.1%-1%. In some embodiments, the proportion of biologically active substance is 1%-20%, 5%-20%, 10%-20%, 15%-20%, or 15%-25%. In some embodiments, the proportion of biologically active substance is less than 0.1%. In some embodiments, the proportion of biologically active substance is at least 25%. In some embodiments, the proportion of biologically active substance is approximately 0.1%, approximately 0.5%, approximately 1%, approximately 2%, approximately 3%, approximately 4%, approximately 5%, approximately 6%, approximately 7%, approximately 8%. , About 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, or more.

Related biologically active substances can be obtained or produced by any available method or manner. The biologically active material may contain or be modified to contain one or more moieties intended to utilize or transport the nanoparticles of the present invention. Such modifications should not interfere with the activity of the biologically active substance. In some embodiments, the modification is in It can be selectively removed in vivo. For example, biologically active substances may be provided in a “pro” form that can be sensibly labeled or converted or transformed into an active form after delivery.

In some embodiments, the biologically active substance is a small molecule and/or organic compound that has pharmaceutically activity. In some embodiments, the biologically active substance is a drug of clinical use. In some embodiments, the drug is an antibiotic, an anti-viral agent, an anesthetic agent, an anticoagulant agent, an anticancer agent, an enzyme inhibitor, a steroid agent, an anti-inflammatory agent, an anti-neoplastic agent, an antigen, a vaccine, an antibody, a decongestant, an antihypertensive agent, a sedative agent, a contraceptive agent, a progesterone agent. , Anticholinergic agents, anesthetics, antidepressants, antipsychotics, beta-adrenergic blockers, diuretics, cardiovascular activators, vasoactive agents, and non-steroidal anti-inflammatory drugs. Of particular interest are biologically active substances suitable for transdermal administration.

The biologically active substance to be delivered can also be a mixture of pharmaceutically active substances. For example, local anesthetics may be delivered in combination with anti-inflammatory agents such as steroids. Local anesthetics may also be administered with vasoactive substances such as epinephrine. For example, antibiotics may be combined with inhibitors of enzymes that inactivate antibiotics commonly produced by bacteria (eg, penicillin and clavulanic acid).

In some embodiments, the biologically active substance is a diagnostic substance. In some embodiments, the diagnostic substance includes a gas; Positron emission tomography (PET); Computed tomography (CAT), single photon emission computerized tomography (single photon emission computerized tomography), x-rays, fluoroscopy, magnetic resonance imaging (magnetic resonance imaging) and commercially available imaging materials and flavoring agents used. Examples of materials suitable as contrast agents used in MRI include gadolinium chelate, iron, manganese, magnesium, copper, and chromium. Materials useful for CAT and x-ray imaging include iodine-based materials.

In some embodiments, the biologically active substance is a prophylactic agent. In some embodiments, the prophylactic agent includes a vaccine. Vaccines may include isolated proteins or peptides, inactivated organisms and viruses, dead organisms and viruses, genetically modified organisms or viruses, and cell extracts. The prophylactic agent may be a combination of interleukin, interferon, cytokine, cholera toxin, alum, and Freund's adjuvant. Preventive drugs include Streptococcus pneumoniae (Streptococccus pnuemoniae), Haemophilus influenza (Haemophilus influenzae), Staphylococcus aureus (Staphylococcus aureus), Strentococcus pyrogen (Streptococcus pyrogenes), Corynebacterium diphtheria (Corynebacterium diphtheriae), Listeria monopsychogen (Listeria monocytogenes), Bacillus anthraces (Bacillus anthracis), Clostridium Tetani (Clostridium tetani), Clostridium botlinium (Clostridium botulinum), Clostridium perfringen (Clostridium perfringens), Niseria Meningitidis (Neisseria meningitidis), Niseria Gonohoe (Neisseria gonorrhoeae), Streptococcus mutans (Streptococcus mutans), Pseudomonas Eruginosa (Pseudomonas aeruginosa), Salmonella Tipi (Salmonella typhi), Haemophilus parainfluenza (Haemophilus parainfluenzae), Bordetella pertussis (Bordetella pertussis), Francisella Turalensis (Francisella tularensis), Ersinia Festival (Yersinia pestis), Vibrio Cholera (Vibrio cholerae), Legionella pneumophilia (Legionella pneumophila), Mycobacterium tuberculosis (Mycobacterium tuberculosis), Mycobacterium repre (Mycobacterium leprae), Treponema Palidum (Treponema pallidum), Leptospirosis interogans (Leptospirosis interrogans), Borreia Burgdorferi (Borrelia burgdorferi), Campylobacter jejuni (Camphylobacter jejuni), and antigens of organisms such as the like; Smallpox, influenza A and B, respiratory syncytial virus, parainfluenza, measles, HIV, varicella-zoster, herpes simplex 1 and 2, cytomegalovirus Megalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps virus, rabies, rubella, coxsackieviruses, equine encephalitis encephalitis), Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, E viruses and antigens of similar viruses; Antigens of fungi, protozoa, and Cryptococcus neoromance (Cryptococcus neoformans), histoplasma capsule ratum (Histoplasma capsulatum), Candida albicans (Candida albicans), Candida Tropicalis (Candida tropicalis), Nocardia astroid (Nocardia asteroides), Rickettsia Rickettsia (Rickettsia ricketsii), Rickettsia Typi (Rickettsia typhi), Mycoplasma New Monie (Mycoplasma pneumoniae), Chlamydial Shitachi (Chlamydial psittaci), Chlamydial Trachomatis (Chlamydial trachomatis), Plasmodium Palsipa Room (Plasmodium falciparum), Trypanozoma Burusei (Trypanosoma brucei), Entamoeva Histolytica (Entamoeba histolytica), Toxoplasma Gondi (Toxoplasma gondii), Trichomonas Vaginalis (Trichomonas vaginalis), Shistozoma Mansoni (Schistosoma mansoni) And similar parasite antigens. These antigens may be intact dead organisms, peptides, proteins, glycoproteins, carbohydrates, or complex forms thereof.

In some embodiments, the biologically active substance may be a protein. As used herein, “protein” and “peptide” are compatible. In certain embodiments, the peptide ranges from about 5 to about 40, about 10 to about 35, about 15 to about 30, or about 20 to about 25 amino acids in size. Peptide panels composed of a variety of sequences may be used to provide a maximally diverse panel of random sequences and/or peptides.

In some embodiments, the biologically active substance may be an antibody. In some embodiments, antibodies include, but are not limited to, polyclonal, monoclonal, chimeric (eg, humanized), single chain (recombinant) antibodies. In some embodiments, antibodies reduce effector function and/or bispecific molecules. In some embodiments, antibodies may also include Fab fragments and/or fragments made by Fab expression libraries.

In some embodiments, the biologically active substance may be a nucleic acid. In some embodiments, the oligonucleotide consists of DNA, RNA, chimeric mixtures, derivatives, specific portions and/or modified versions thereof. The oligonucleotide of the present invention may be single-stranded and/or double-stranded.

Oligonucleotides can also be modified in the base moiety, sugar moiety and/or phosphate backbone to improve the stability and hybridization of the molecule.

In certain embodiments, the nucleic acid consists of an antisense molecule that binds to the translation initiation site and/or splice junction. Antisense oligonucleotides will bind to the target mRNA or interfere with translation. Alternatively or additionally, antisense oligonucleotides may also bind to the DNA of a target gene, such as a regulatory element.

In some embodiments, the nucleic acid comprises a ribozyme designed to catalytically cleave a target mRNA transcript that can be used to disrupt translation of the target mRNA or effect expression of the target (PCT publication WO 90/1 1364; and Sarver et al., 1990, Science 247:1222; these documents are incorporated by reference).

Alternatively or additionally, endogenous target gene expression can be reduced by targeting a deoxyribonucleotide sequence complementary to the regulatory portion of the target gene to form a triple helix structure that interferes with the transcription of the target gene in the target muscle cell in the body. see generally, Helene, 1991, Anticancer Drug Des. 6:569; Helene et al., 1992, Ann, N.Y. Acad. Sci. 660:27; and Maher, 1992, Bioassays 14:807; All documents are attached as reference materials).

In some embodiments, the biologically active substance is a nutritional substance. In some embodiments, the nutritional substance provides basic nutritional value. In some embodiments, the nutritional substance provides a health or medical benefit. In some embodiments, the nutritional substance is a dietary supplement.

In some embodiments, the nutritional substance is a vitamin. In some embodiments, the vitamin is one or more vitamin A (retinoids), vitamin Bl (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyroxidone), vitamin B7. (biotin), vitamin B9 (folic acid), vitamin B12 (cyanocobalamin), vitamin C (ascorbic acid), vitamin D, vitamin E, or vitamin K.

In some embodiments, the nutritional substance is a mineral. In some embodiments, the mineral is one or more of bismuth, boron, calcium, chlorine, chromium, cobalt, copper, florin, iodine, magnesium, manganese, molybdenum, nickel, phosphorus, potassium, rubidium, selenium. , Silicon, sodium, strontium, sulfur, tellurinum, titanium, tungsten, vanadium or zinc.

In some embodiments, the nutritional substance is an essential amino acid. In some embodiments, the amino acid is one or more of arginine, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, or valine.

In some embodiments, the nutritional substance is fatty acids and/or omega-3 fatty acids (e.g., DHA or ARA), fruit and vegetable extracts, lutein, phosphatidylserine, lipoic acid, melatonin, glucosamine, chondroitin, aloe vera, guggul ), green tea, lycopene, whole foods, food additives, herbs, phytonutrients, antioxidants, flavonoids in fruits, evening primrose oil, flaxseed, fish and marine animal oils (e.g. cod liver oil ) And protobiotics may be included. In some embodiments, nutritional substances may also include bio-engineered foods (also referred to as functional foods “pharmafoods”) that have been genetically engineered to have the desired properties.

Exemplary nutritional substances and dietary supplements are described in Roberts et al, Nutriceuticals: The Complete Encyclopedia of Supplements, Herbs, Vitamins, and Healing Foods, American Nutriceutical Association, 2001; incorporated herein by reference. Nutritional substances and dietary supplements are also known in Physicians' Desk Reference for Nutritional Supplements, 1st Ed., 2001, and Physicians' Desk Reference for Herbal Medicines, 1st Ed., 2001 (these documents are incorporated by reference).

In some embodiments, the nanoparticles of the present invention loaded with nutritional substances may be incorporated into food products. For example, nutrient-loaded nanoparticles may be dissolved in liquids such as beverages.

In some embodiments, the biologically active substance is a cosmetic and/or skin substance. In some embodiments, cosmetic and/or skin substances include vitamins and derivatives thereof (e.g., vitamin E and esters thereof, vitamin C and esters thereof, vitamin B, vitamin A alcohol or retinol and esters thereof), Provitamins (e.g. panthenol, niacinamide or ergalciferol), antioxidants, phenolic compounds (e.g. benzoyl peroxide), essential oils, humectants, sunscreens, moisturizers, proteins, ceramides and pseudos. Ceramide and the like are included.

In some embodiments, the biologically active material may also include one or more botlinium toxin peptides or protein complexes. In some embodiments, the botlinium toxin can be one or more botlinium toxin serotypes A, B, C ₁ , C ₂ , D, E, F, or G. In some embodiments, the botlinium toxin can be an isolated and/or purified toxin. In some embodiments, the botlinium toxin can be a partially isolated and/or partially purified botlinium toxin. In some embodiments, the botlinium toxin may be a native botlinium complex. In some embodiments, the botlinium toxin may be associated with a non-toxin protein. In some embodiments, the botlinium toxin may be a recombinantly made botlinium toxin.

One of skill in the art will appreciate that this is provided as an example and is not an exhaustive list of biologically active substances. Any biologically active material may be entrapped within the nanoparticles or bound to the surface.

Delayed release substances

In some embodiments of the invention, particularly those comprising one or more biologically active substances (e.g., unmodified peptides), the nanoparticle compositions of the invention further comprise one or more release-delaying components. So that the release of the active substance can be controlled. Any release-delaying component known in the art is suitable for use in making the nanoparticles of the present invention. In some embodiments, the release delaying components are hydrophilic and/or hydrophobic polymers. Release-delaying components include, for example, cellulose or derivatives thereof, acrylic polymers, ester polymers, vinyl-pyrrolidone-based polymers, gums, other natural polymers, and/or composites thereof.

In some embodiments, the release delaying component is cellulose or a derivative thereof. In certain embodiments, cellulose or a derivative thereof is one or more of hydroxypropyl methylcellulose, methylcellulose, carbooxymethylcellulose, carbooxymethylcellulose sodium, hydroxypropyl ethylcellulose, hydroxyethylcellulose, and hydroxy It is propyl cellulose. In certain embodiments, the cellulose or a derivative thereof is methylcellulose or a derivative thereof. In certain embodiments, the cellulose or a derivative thereof is hydroxypropyl methylcellulose (HPMC). One of skill in the art will recognize that other polymers can be used, including alkyl cellulose polymers.

In some embodiments, the release-delaying component is an acrylic polymer. In certain embodiments, acrylic polymers include, for example, acrylic acid and methacrylic acid co-polymers, methyl methacrylate co-polymers, ethoxyethyl methacrylate, cyanoethyl methacrylate, aminoalkyl methacrylate co-polymers. -Polymer, poly(acrylic acid), poly(methacrylic acid), methacrylic acid alkylamide co-polymer, poly(methyl methacrylate), poly(methacrylic acid anhydride), methyl methacrylate, polymethacrylate , Poly(methyl methacrylate) co-polymer, polyacrylamide, aminoalkyl methacrylate co-polymer, glycidyl methacrylate co-polymer, and composites comprising one or more of the aforementioned polymers. The acrylic polymer may be composed of a copolymer of fully polymerized acrylic and methacrylic acid esters with a low content of tetravalent ammonium groups.

In some embodiments, the release-delaying component is a polyester. In some embodiments, polyesters include polyalkylene glycol, poly(glycolide-co-lactide), pegylated poly(latic-co-glycolic acid), poly(lactic acid), pegylated poly(lactic acid), Poly(glycolic acid), pegylated poly(glycolic acid), polylactic acid and copolymers of polyglycolic acid and derivatives thereof. In some embodiments, polyesters include, for example, polyanhydride, poly(ortho ester) pegylated poly(ortho ester), poly(caprolactone), pegylated poly(caprolactone), polylysine , Pegylated polylysine, poly(ethylene imine), pegylated poly(ethylene imine), and derivatives thereof. In some embodiments, polyesters include, for example, polycaprolactone, poly(L-lactide-co-L-lysine), poly(serine ester), poly(4-hydroxy-L-proline ester), poly [α-(4-aminobutyl)-L-glycolic acid] and derivatives thereof.

In some embodiments, the release-delaying component is a cross-linked polymer of poly(vinyl-pyrrolidone). In some embodiments, the polymer is crospovidone. In some embodiments, the polymer is uncrosslinked poly(vinyl pyrrolidone). In some embodiments, the polymer is povidone.

In some embodiments, the delayed-release component can be a natural polymer. In some embodiments, the natural polymer is a gum such as xanthan gum, alginic acid, karaya gum, alginate sodium and/or locust bean gum. In some embodiments, the natural polymer may be a protein (eg, albumin), a lipid, a nucleic acid, or a carbohite. The release-delaying component is polyester. In some embodiments, polyesters include polyalkylene glycol, poly(glycolide-co-lactide), pegylated poly(latic-co-glycolic acid), poly(lactic acid), pegylated poly(lactic acid), It is a co-polymer of poly(glycolic acid), pegylated poly(glycolic acid), polylactic acid and polyglycolic acid. In some embodiments, polyesters include, for example, polyanhydride, poly(ortho ester) pegylated poly(ortho ester), poly(caprolactone), pegylated poly(caprolactone), polylysine , Pegylated polylysine, poly(ethylene imine), pegylated poly(ethylene imine), and derivatives thereof. In some embodiments, polyesters include, for example, polycaprolactone, poly(L-lactide-co-L-lysine), poly(serine ester), poly(4-hydroxy-L-proline ester), poly [α-(4-aminobutyl)-L-glycolic acid] and derivatives thereof.

In some embodiments, the release delaying component is a cross-linked polymer of poly(vinyl-pyrrolidone). In some embodiments, the polymer is crospovidone. In some embodiments, the polymer is uncrosslinked poly(vinyl pyrrolidone). In some embodiments, the polymer is povidone. In some embodiments, the delayed-release component can be a natural polymer. In some embodiments, the natural polymer is a gum such as xanthan gum, alginic acid, karaya gum, alginate sodium and/or locust bean gum. In some embodiments, the natural polymer may be a protein (eg, albumin), a lipid, a nucleic acid, or a carbohite.

Formulation material ( Formulating Agent )

The nanoparticle composition of the present invention can be made to be administered to a subject. In certain embodiments, the nanoparticle composition of the present invention can be administered to the skin and transdermally delivered to a subject. For example, the nanoparticle composition of the present invention may be formulated into a cosmetic or other formulation for topical application.

Human skin consists of the dermis and epidermis. The epidermis consists of several layers of tissue, such as the stratum corneum, the stratum lucidum, the stratum granulosum, the stratum spinosum, and the stratum basale (inward to the outer surface of the skin). ). The stratum corneum is generally the most important obstacle for transdermal delivery of drugs, and perhaps the largest, especially for the delivery of unmodified peptides. The stratum corneum is generally about 10-15 μm thick and consists of flat keratised cells (keratised cells) arranged in several layers. The intercellular space between keratinocytes is filled with lipid structures and will play an important role when substances penetrate through the skin (Bauerova et ah, 2001, European Journal of Drug Metabolism and Pharmacokinetics, 26:85; attached as a reference). .

The rest of the dermis under the stratum corneum is approximately 150 μm thick. The dermis is about 1 mm-2 mm thick and is located below the epidermis. The dermis is nerved by various capillaries and neural processes.

In general, attempts for transdermal administration of drugs have focused on increasing the permeability of the stratum corneum. Some attempts have included the use of chemical enhancers that increase the penetration of molecules through the skin. Some attempts include the use of mechanical devices that bypass or remove portions of the stratum corneum. It also includes the use of ultrasound or iontophoresis to effect penetration of the drug through the skin. In most cases, the goal is to move a pharmaceutical substance, usually small molecule, through the skin to a capillary bed in the dermis, where the substance systematically binds to the subject to obtain a therapeutic effect.

The present invention provides, among others, a method of transdermally administering an unmodified peptide without the use of a friction agent or other bursting agent (chemical, mechanical, electrical, magnetic, etc.). In the present invention, it was surprisingly discovered that when botulinum toxin is combined with the nanoparticle composition of the present invention, it is effectively transmitted transdermally without penetration or rupture of the stratum corneum. The use of such materials or steps with the botlinium nanoparticle compositions of the present invention is not necessarily excluded or required in all embodiments of the present invention.

The present invention provides a method of administering an unmodified peptide through topical provision of the nanoparticle composition of the present invention. In some embodiments, the nanoparticle composition of the present invention is provided directly through the skin and allowed to be absorbed through the epidermal layer. In some embodiments, the nanoparticle composition is capable of penetrating the stratum corneum, dermal pores, and/or the upper layer of teeth, including the dermal gland, without the use of chemical or mechanical skin penetration enhancers or other substances that cause friction.

Those skilled in the art will appreciate that the composition of the present invention for topical administration is a skin softener, a nourishing lotion type emulsion, a cleansing lotion, a cleansing cream, a skin milk, an emollient lotion, a massage cream, an emollient cream, a makeup base, a lipstick, a face pack, or a face. It will be appreciated that it may be provided in the form of a skin composition such as a gel, shampoo, conditioner, body-cleanser, hair-tonic, cleaner product such as soap, lotion, study, gel, cream, patch or spray.

The formulation of the nanoparticle composition of the present invention may include one or more excipients, such as a filler, a sequestering agent, an emollient, a color developing material (eg, dyes and pigments), and a fragrance.

In some embodiments, the nanoparticle composition of the present invention may be formulated as a cream. “Cream” refers to a composition that can be applied, and is generally a form for applying to the skin. Creams generally contain oil and/or fatty acid-based media. The cream according to the present invention contains nanoparticles, and substantially completely penetrates (nanoparticles) through the skin when administered topically. Such creams can also act as carriers of bound substances (eg, botulinum toxin).

One of skill in the art will recognize that the nanoparticle compositions of the present invention can be incorporated into devices such as patches.

Transdermal patch structures are variously known in the art; One of skill in the art will recognize that the nanoparticle compositions of the present invention can be directly bonded to any of such a variety of structures. In some embodiments, the transdermal patch may further include multiple needles extending from one side of the patch when provided to the skin, with the needles extending from the patch being fired through the stratum corneum of the skin. In some embodiments, the needles do not rupture the blood vessel.

In some embodiments of the invention, the nanoparticle composition is provided as a depot to the patch such that the pressure applied to the patch directs the unmodified peptide out of the patch (optionally through needles) and through the stratum corneum.

In some embodiments of the present invention, the transdermal patch includes an adhesive. Examples of adhesives are well known in the art (e.g., U.S. Patents 296,006; 6,010,715; 5,591,767; 5,008,110; 5,683,712; 5,948,433; 5,965,154; all incorporated by reference). Adhesive patches generally have a characteristic configuration with an adhesive layer to adhere to the skin, a depot or reservoir to hold the drug, and an outer surface that prevents leakage of the drug from the depot. The outer surface of the patch is generally non-adhesive.

Those skilled in the art will understand that the transdermal patch is only one example of administering the nanoparticle composition of the present invention. As another example, unnecessary numbness of the finger can be avoided by using a device capable of providing the composition to the finger of a person without being primarily different from the composition. Suitable devices include spatulas, swabs, needleless syringes, and adhesive patches. The device should be inserted into the container containing the composition using a spatula or cotton bottle or similar. When using a syringe, fill the syringe with the composition and use it. The composition is applied topically by spreading it with a spatula or cotton swab, and may be released through a syringe into the skin.

In many embodiments of the present invention, it may be desirable to restrict delivery of the unmodified peptide to only the desired delivery site. In some embodiments, such limited delivery can be effected using the nanoparticle composition of the present invention in a device that allows delivery of the composition to only a desired target area of the skin without delivering the composition to other areas of the skin. Obviously, a transdermal patch can be used for this purpose. Alternatively or additionally, if the modified peptide is provided topically to a selected site, the other site must be masked or pretreated or otherwise protected from exposure.

<Example>

The following examples are intended to describe specific examples considered in the present invention. This embodiment is not limited in any form.

Example 1: Pentapeptide nanoparticle composition.

This example is KTTKS (SEQ ID NO.: 1), known to have biological activity in skin structure (Katayama et al, supra). It provides a peptide nanoparticle composition composed of a nanoemulsion containing pentapeptide.

The pentapeptide nanoemulsion formulation was prepared as follows:

• 800mg soybean oil, 800mg Tween 80 are stirred in a sterile vial for 5 minutes;

-8.4 ml water and 0.0001 g KTTTS (SEQ ID NO.: 1) were added and stirred for 20 minutes;

Samples were homogenized for 1 minute;

-The sample was stirred for 20 minutes;

Samples were microfluidized once at 23,000 psi.

The resulting pentapeptide nanoemulsion was evaluated using a Malvern Nano S particle classifier capable of classifying particle sizes of about 0.6 nm-6000 nm. The pentapeptide nanoemulsion preparation has two particle size peaks with an average particle size of 106 nm (Table 2).

Size range Particle ratio 10-20nm 1.3% 21-100nm 30.2% 101-120nm 10.4% 121-150nm 22.4% 151-200nm 19.3% 201-300nm 14.7% 301-400nm 1.7% synthesis 100.0%

Example 2: pentapeptide nanoparticles Preparation And biological effects Transdermal Penetration

In this example, KTTKS (SEQ ID NO.: 1), known to have biological activity in skin structure (Katayama et al, supra). It provides a peptide nanoparticle composed of a nanoemulsion containing pentapeptide. In this example, in the case of KTTKS (SEQ ID NO.: 1), the biological effect when the peptide nanoparticles are transdermally provided to the skin will be described.

Materials and methods

The pentapeptide nanoemulsion formulation was prepared as follows:

5.6 g Labrafac WL 1349 oil and 5.6 g Tween 80 are stirred in a sterile vial for 5 minutes;

• 58.8g reagent grade water is placed in a separate beaker; 0.010g peptide KTTTS (SEQ ID NO.: 1) was added to water and stirred for 20 minutes.

The contents of the first beaker were put into the beaker contents (water and peptide) and stirred for 20 minutes; And

● The entire sample was microfluidized once at 23,000 psi.

The resulting pentapeptide nanoemulsion was evaluated using a Malvern Nano S particle classifier capable of classifying particle sizes of about 0.6 nm-6000 nm. The pentapeptide nanoemulsion preparation has an average particle size of 114.4 nm. Approximately 95% of the particles are less than 130 nm in size.

Pentapeptide nanoemulsion is mixed with an equal amount of skin cream (Base PCCA Vanishing Cream Light) and vortexed until a homogeneous cream is obtained to make a “treatment cream”.

The control cream ("Control Cream") is made in the same way, but no peptides are added in the process.

Buy 10 Swiss Webster mice (20 g worms each). Upon arrival, all animals were allowed to acclimate in their cages for 1 week (divided into 5 cages as described below) and provided with standard cage bedding and Purina 5001 chow. After 1 week it was treated as follows;

Processing process

Group 1 (Control): Every day for 8 weeks, put on gloves on the backs of 5 rats and apply 75 µl of “Control Cream” until no cream is visible. The backs of mice are dehaired with an electric razor two days before the first treatment, and then removed at intervals of one week.

Group 2 (treatment group): Every day for 8 weeks, put on gloves on the backs of 5 rats and apply 75 µl of “treatment cream” until no cream is visible. The backs of mice are dehaired with an electric razor two days before the first treatment, and then removed at intervals of one week. The backs of mice are dehaired with an electric razor two days before the first treatment, and then removed at intervals of one week.

evaluation

The skin of the back of each mouse was treated with a control cream or a treatment cream, and was stained with Masson's Trichrome tissue. The color intensity was evaluated at 400x magnification using a degree of tissue coloration of 1 to 4; 1 = Almost no coloration. Collagen fibrils are very slender; 2 = minimum pigmentation, minimum collagen fiber width; 3 = medium pigmentation, medium fiber width; 4 = strong pigmentation and wide fiber

result

Histological evaluation

The average histological evaluation of skin tissue stained with Masson's Trichrome staining was 2.33 in the control group, and 3.67 in the treatment group. This indicates that the collagen staining intensity was increased by 57% in the treatment group compared to the control group. 1 is a photomicrograph of a skin tissue sample of each of the control group and the treatment group.

Evaluation of skin thickness effect

The skin thickness was determined by examining the tissue fragments of the skin such as mice placed on a finely cut glass slide using the Skin Layer Thickness test to determine the thickness (mm) of each skin layer.

Extracellular Matrix production evaluation

Collagen is a major component of the extracellular matrix. Collagen content was assessed by using two distinct tissue stains (Picro Sirius Red and Pterocarpus Osun) to perform two separate tests for collagen on tissue fragments of the skin such as mice placed on finely chopped glass slides.

Collagen content was evaluated using Western Blot technology, which detects the content of hydroxyproline in skin such as homogenized mice. Hydroxyproline content refers to collagen content.

conclusion

Histological evaluation in the results showed that the treatment group had statistically more collagen than the control group. These data show that topical pentapeptide nanoemulsion formulations have measurable biological effects on the skin when compared to control creams without such peptides. Previous studies have suggested that peptides cannot penetrate native skin without chemical modification (Katayama et al, supra). Thus, these data show that the nanoemulsion formulation of the present invention allows the unmodified peptide to penetrate, thereby increasing collagen production in the skin, resulting in results consistent with the biological action of known peptides to increase skin thickness. .

These results are expected that, on average, the treatment group had statistically thicker skin than the control group. These results show that, as confirmed by two histological staining and Western Blot measurements of hydroxyproline, statistically more collagen was retained in the treatment group than in the control group.

Example 3; Skin thickening and stimulating effects of extracellular matrix in mice through transdermal provision of peptide nanoparticles; Effects of varying the concentration of peptides in nanoparticles

In this example, the effect of providing the peptide nanoparticles transdermally to the skin and varying the concentration of the peptides in the nanoparticles in terms of biological effects will be described.

Materials and methods

The experiment of Example 3 was repeated, except that the peptide concentration in the treatment cream was increased by 10 or decreased by 10.

Results and conclusion

The results show that increasing the peptide concentration showed that the skin of mice treated with the increased peptide concentration was statistically thicker than that of the mice treated with the reduced peptide concentration. As confirmed by the two histological staining and Western Blot measurements of hydroxyproline, the results showed that increasing the peptide concentration on average resulted in the skin of mice treated with the increased concentration of peptide being treated with the peptide with a statistically reduced concentration. It has been shown to have more collagen than mice.

In summary, with these data, it can be said that the effect of the peptide nanoemulsion on the skin depends on the concentration of the peptide contained therein.

Example 4: skin line ) administration of pentapeptide nanoparticles to humans

This example demonstrates the biological effects of transdermal administration of peptide nanoparticles.

Materials and methods

The pentapeptide nanoemulsion prepared according to Example 1 or 2 is mixed with an equal amount of skin cream (Base PCCA Vanishing Cream Light), and vortexed until a homogeneous cream is obtained to obtain a "treatment cream".

Mixing the same amount of water and the same amount of pentapeptide as in Example 1, and mixing with the same amount of the same skin cream used when making the treatment cream, to make a "non-nano-treated cream".

The “control cream” is made by vortexing the same amount of the material in Examples 1 or 2 and the same skin cream used to make the treatment cream.

Thirty healthy subjects with facial wrinkles (mainly observed in people with sun-damaged skin) were enrolled in a double-blind, placebo-controlled, left and right random split-faced study. All subjects receive a 5-stage score rating by an observer who is unaware of the treatment status. The 0-knot is normal skin, and the 5th grade is severe facial wrinkles and lines (mainly around the eyes or around the eyes). Cheek skin tissue is evaluated by pore size (small to large) and softness (soft to coarse/gravel-like). Registration was made only when the initial value was 2.5 or higher. The subject's face will be photographed using standardized observation view, distance, and light conditions.

Processing process

The patient agrees not to use any facial skin care products except for the control cream twice daily at 12 hour intervals for 3 weeks. After the initial “wash-out” period, each patient is given two tubes labeled “left” and “right” with a unique numeric code for each tube. Each person is instructed to apply a tube labeled “right” on the right side of the face and a tube labeled “left” on the left side twice a day at 12-hour intervals. You are instructed to apply a pea-sized cream (about 0.4g) to each side of the face. You are also instructed not to use other facial care products. In the case of 10 individuals (control group), the right tube contains the control cream and the left tube contains the control cream. In the case of 10 subjects (non-nano treatment group), the right tube contained the control cream and the left tube contained the non-nano treatment cream. In the case of 10 individuals (nano treatment group), the control cream was contained in the right tube and the nano treatment cream was contained in the left tube.

evaluation

Subjects begin the course of treatment after the wish-out period and are observed and photographed at 4, 8 and 12 weeks. In addition, an observer who does not know about the treatment of the individual and the individual themselves evaluate the skin texture on the left and right sides of the face by the above-described numerical values.

Results and conclusion

In the results, on average, the nano-treatment group had a statistically greater difference in facial texture (improvement of skin appearance) than the difference between the left and right values observed in the major group and the non-nano treatment group. In summary, these data are measurable when a topical pentapeptide nanoemulsion formulation is compared to a control cream without pentapeptide and a cream with the same pentapeptide not included in the nanoparticle formulation (non-nano cream). I think it has a degree of cosmetic effect.

Equivalent range

The above is a description of suitable non-limiting embodiments of the invention. Those of skill in the art, using routine experimentation, will recognize or be assured that there are many equivalents to the specific embodiments described herein. Those skilled in the art will appreciate that various changes and modifications to such description will not depart from the scope of the invention as defined by the claims appended hereto.

In the claims, articles ("a," "an," "the") mean one or more unless otherwise stated. In the claims and specification, an “or” between one or more numbers between groups is deemed to be all groups or one or more used. The present invention includes embodiments that provide or use a group's exact value. The invention also includes embodiments in which one or more or all of them can be used in a group configuration. In addition, the present invention also includes one or more limitations, variations, complexes, and the like in the relevant parts of one or more claims or descriptions. For example, any term that is dependent on one term may be modified to include one or more restrictions found in other terms that are dependent on the same term.

In addition, where the claim refers to a composition, also includes methods of using the composition for any of the purposes described herein, including methods of making the composition according to any of the methods described herein. For example, it will be appreciated that any composition of the present invention can be used to prevent the formation, progression or recurrence of adsorption at any location due to any of the causes discussed herein. In addition, it will be appreciated that any composition made according to the method of making the compositions described herein can be used to prevent adsorbent formation, progression or recurrence at any location for any cause discussed herein. In addition, the present invention includes compositions made according to the method of making the compositions described herein.

When elements are provided in the form of a list (Markush group format), each subgroup of each element is described, and it should be understood that arbitrary elements may be omitted from the group. Also, “comprising” is in the open sense to allow additional elements or steps to be included. In general, it can be said that the invention or aspect of the invention is included as a specific element, feature, step, etc. of the invention, or is basically composed of such elements, steps, features, and the like. For similar purposes these embodiments are not specifically presented herein. Each embodiment of the present invention including one or more elements, features, steps, etc. may also provide embodiments consisting of or consisting essentially of these elements, features, steps, and the like.

For ranges, the endpoints are included. Further, unless otherwise stated, the numerical values indicated in the ranges may represent 1/10 of the lower limit unit of this range for any particular value in the ranges mentioned in the different embodiments of the present invention. In addition, unless otherwise stated, numerical values expressed in a range can estimate any sub-range of a given range, and both ends of the sub-range may be expressed as 1/10 the lower limit of the range.

In addition, it should be understood that any particular embodiment of the invention may be explicitly excluded from one or more claims. Any embodiments, elements, features, applications and aspects of the compositions and/or methods of the present invention (e.g., any peptide, any peptide modification, any nanoparticles, any nanoemulsion, any surfactant, any oil, any pre- It should be understood that mix components, any methods of making nanoemulsions, arbitrary treatment methods, etc.) may be explicitly excluded from one or more claims. All embodiments excluding one or more elements, features, purposes or aspects for the purpose of brevity are not presented herein.

SEQUENCE LISTING <110> Edelson, Jonathan Kotyla, Timothy <120> PEPTIDE NANOPARTICLES AND USES THEREFOR <130> 2007339-0016 <140> PCT/US2007/086040 <141> 2007-11-30 <160> 14 <170> PatentIn version 3.5 <210> 1 <211> 5 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 1 Lys Thr Thr Lys Ser 1 5 <210> 2 <211> 10 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 2 Glu Tyr Lys Thr Thr Lys Ser Ser Arg Leu 1 5 10 <210> 3 <211> 8 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 3 Val Ile Glu Tyr Lys Thr Thr Lys 1 5 <210> 4 <211> 4 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 4 Lys Thr Thr Lys One <210> 5 <211> 12 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 5 Gly Lys Thr Val Ile Glu Tyr Lys Thr Thr Lys Ser 1 5 10 <210> 6 <211> 15 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 6 Gly Lys Thr Val Ile Glu Tyr Lys Thr Thr Lys Ser Ser Arg Leu 1 5 10 15 <210> 7 <211> 20 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 7 Trp Gly Lys Thr Val Ile Glu Tyr Lys Thr Thr Lys Ser Ser Arg Leu 1 5 10 15 Pro Ile Ile Asp 20 <210> 8 <211> 20 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 8 Cys Thr Ser His Thr Gly Ala Trp Gly Lys Thr Val Ile Glu Tyr Lys 1 5 10 15 Thr Thr Lys Ser 20 <210> 9 <211> 4 <212> PRT <213> Artificial sequence <220> <223> peptide that may promote extra-cellular matrix production <400> 9 Thr Thr Lys Ser One <210> 10 <211> 6 <212> PRT <213> Artificial sequence <220> <223> peptide that may decrease wrinkles <400> 10 Glu Glu Met Gln Arg Arg 1 5 <210> 11 <211> 6 <212> PRT <213> Artificial sequence <220> <223> peptide that may improve wound healing <400> 11 Val Gly Val Ala Pro Gly 1 5 <210> 12 <211> 6 <212> PRT <213> Artificial sequence <220> <223> peptide that may improve wound healing <400> 12 Tyr Tyr Arg Ala Asp Ala 1 5 <210> 13 <211> 3 <212> PRT <213> Artificial sequence <220> <223> peptide that may improve wound healing <400> 13 Gly His Lys One <210> 14 <211> 14 <212> PRT <213> Artificial sequence <220> <223> peptide that may treat accumulation of excess extracellular matrix <400> 14 Thr Ser Leu Asp Ala Ser Ile Ile Trp Ala Met Met Gln Asn 1 5 10

Claims (1)

Wherein the majority of the particles have a diameter of from 10 to 300 nm and the nanoparticles are selected from the group consisting of EYKTTKSSRL, GKTVIEYKTTKS, GKTVIEYKTTKSSRL, WGKTVIEYKTTKSSRLPIID, and CTSHTGAWGKTVIEYKTTKS, and the unmodified peptide of the amino acid sequence selected from the group consisting of one species Or more, and has a biological activity on skin, subcutaneous tissue or adjacent muscle.

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